Extraction method of Tremella polysaccharide

Abstract

The present invention relates to the technical field of natural active product extraction, and in particular to a method for extracting tremella polysaccharide; the raw material for extracting polysaccharide is directly fresh tremella, and no pre-treatment steps such as drying and crushing are required, so as to improve the extraction efficiency; the fresh tremella is first soaked in low-temperature clean water, so that there is sufficient water inside and outside the fresh tremella cells; then the fresh tremella is quickly frozen, so that ice crystals are formed inside and outside the cells during the quick freezing process, so that the cell membrane and cell wall of the fresh tremella are squeezed and broken, and then microwave radiation is used to cause the cell membrane and cell wall to be further broken, and then high-voltage pulse electric field treatment is used, and short pulses of high voltage can be used to further accelerate the release of polysaccharide components. The tremella cells are completely broken by physical means, so that the polysaccharide components are accelerated to dissolve, and the whole process is completed under low temperature conditions, and high-temperature enzyme inactivation or high-temperature extraction is not required, so that the heat-sensitive substances are guaranteed not to be degraded, thereby ensuring that the structure of the tremella polysaccharide is not destroyed.

Description

Technical Field

[0001] The invention relates to the technical field of natural active product extraction, in particular to a method for extracting tremella polysaccharide. Background Art

[0002] Tremella fuciformis Berk, also known as white fungus, snow fungus, and tremella, originated in Tongjiang, Sichuan, and is widely distributed. It has the effects of nourishing yin and moistening the lungs, nourishing the stomach and promoting fluid production. It is effective in treating lung heat cough, gastritis, facial spots, cancer tumors and other diseases. Most of its biological activities are related to tremella polysaccharides. It is a traditional fungal food medicine in my country.

[0003] Tremella fuciformis polysaccharides (TFPS) has multiple functional effects such as regulating immunity, lowering blood sugar and blood lipids, anti-radiation, anti-tumor, anti-aging and anti-oxidation; it affects the body's immunity by regulating humoral immunity, cellular immunity and immune factors; it promotes the excretion of cholesterol by adsorbing and binding lipids in the blood, blocking its enterohepatic circulation to achieve the effect of lowering blood lipids; and it has anti-tumor effects as an interferon inducer, activating the body's phagocytes, accelerating their proliferation and increasing their phagocytic ability; it also has anti-aging and antioxidant effects, can effectively protect the skin and remove free radicals in the body.

[0004] Currently, the extraction methods of Tremella polysaccharides include water extraction, enzyme extraction and alkaline extraction.

[0005] The disadvantages of the water extraction method are that it is time-consuming and has a low extraction rate. In addition, in order to increase the extraction rate of Tremella fuciformis polysaccharides, it is usually necessary to carry out the process at high temperatures, which can easily lead to the destruction of the spatial structure of Tremella fuciformis polysaccharides.

[0006] The advantage of alkaline extraction is that it can improve the extraction rate compared to water extraction, but some glucose chains of polysaccharides will degrade when the alkalinity is strong.

[0007] The enzyme extraction method generally uses a complex enzyme such as pectinase, protease, cellulase, etc. to treat Tremella fuciformis, which can accelerate the release and extraction of Tremella fuciformis polysaccharides to a certain extent, but the introduction of the complex enzyme requires subsequent enzyme inactivation treatment.

[0008] Therefore, there is an urgent need to improve the extraction method of Tremella fuciformis polysaccharide so as to increase the extraction rate and efficiency of Tremella fuciformis polysaccharide while avoiding the destruction of the spatial structure of Tremella fuciformis polysaccharide. Summary of the invention

[0009] The technical problem to be solved by the present invention is that it is urgent to improve the extraction method of tremella polysaccharide, and to improve the extraction rate and extraction efficiency of tremella polysaccharide while avoiding the destruction of the spatial structure of tremella polysaccharide.

[0010] In order to solve the above technical problems, the technical solution adopted by the present invention is:

[0011] The present invention relates to a method for extracting Tremella polysaccharide, comprising the following steps:

[0012] Step 1: Wash the fresh Tremella fuciformis, completely immerse it in clean water at 10-30°C for 2-6 hours, and then quickly freeze the soaked fresh Tremella fuciformis;

[0013] Step 2: Mixing the quick-frozen fresh Tremella fuciformis with ice cubes in a weight ratio of 1:18-22, and subjecting the mixture to microwave radiation treatment, wherein the microwave radiation intensity of the microwave radiation treatment is 4000-5000 MHz and the treatment time is 3-5 minutes, thereby obtaining Tremella fuciformis water;

[0014] Step 3: placing the Tremella water in a bipolar square wave high voltage pulse electric field, wherein the pulse electric field strength is 30-40 kV / cm, the frequency is 1100-1200 Hz, and the treatment time is 2-3 hours, to separate the Tremella from the water, and the obtained water is the extract;

[0015] Step 4: Concentrate the extract to one-fourth to one-fifth of its volume, add 95% ethanol, the volume ratio of ethanol to the extract being 3-4:1, and separate the precipitate, which is the crude Tremella polysaccharide.

[0016] Furthermore, in the above-mentioned method for extracting Tremella polysaccharide, after step 4, it also includes step 5: washing the obtained crude Tremella polysaccharide with anhydrous ethanol, anhydrous ether and acetone in sequence, drying and crushing to obtain Tremella polysaccharide.

[0017] Furthermore, in the above-mentioned method for extracting Tremella polysaccharide, in step 3, the temperature of the Tremella water is maintained at 30-35°C.

[0018] Furthermore, in the above-mentioned Tremella polysaccharide extraction method, in step 1, the temperature of the clean water is 20-23° C., and the soaking time is 3-4 hours.

[0019] Furthermore, in the above-mentioned method for extracting Tremella polysaccharide, in the step 2, the weight ratio of the quick-frozen fresh Tremella to ice cubes is 1:20, and the microwave radiation intensity of the microwave radiation treatment is 4500 MHz.

[0020] Furthermore, in the above-mentioned Tremella polysaccharide extraction method, in step 3, the pulse electric field strength is 35 kV / cm, the frequency is 1100 Hz, and the processing time is 2.5 h.

[0021] The beneficial effects of the present invention are as follows: in the tremella polysaccharide extraction method of the present invention, the raw material for extracting the polysaccharide is directly fresh tremella, and no pre-treatment steps such as drying and crushing are required, thereby improving the extraction efficiency and effectively avoiding the problem of reduced extraction rate caused by material loss in the crushing process. In the present scheme, the fresh tremella is firstly soaked in low-temperature clean water to allow sufficient water inside and outside the fresh tremella cells, and then the fresh tremella is quickly frozen. Since the density of ice is lower than that of liquid water, ice crystals are formed inside and outside the cells during the quick freezing process, so that the cell membrane and cell wall of the fresh tremella are squeezed and ruptured, and then microwave radiation is used to make the polar substances in the cells obtain a large amount of heat, causing the temperature in the cells to rise, and the pressure generated by the vaporization of liquid water causes the cell membrane and cell wall to further rupture, so that each cell can form tiny holes, accelerating the release of polysaccharide components in the cells into the water, and after the microwave treatment, high-voltage pulse electric field treatment is used, and the release of polysaccharide components can be further accelerated by using short pulses of high voltage. In the above extraction method, there is no need to introduce complex enzymes to rupture the cell membrane and cell wall. The Tremella cells are ruptured completely by physical means to accelerate the dissolution of the polysaccharide components. The whole process is completed under low temperature conditions, and there is no need for high-temperature enzyme inactivation or high-temperature extraction, thereby ensuring that heat-sensitive substances are not degraded, thereby ensuring that the structure of Tremella polysaccharides is not destroyed. DETAILED DESCRIPTION

[0022] In order to explain the technical content, achieved objectives and effects of the present invention in detail, the following is an explanation in conjunction with the implementation modes.

[0023] The method for extracting Tremella polysaccharide is characterized by comprising the following steps:

[0024] Step 1: Wash the fresh Tremella fuciformis, completely immerse it in clean water at 10-30°C for 2-6 hours, and then quickly freeze the soaked fresh Tremella fuciformis;

[0025] Step 2: Mixing the quick-frozen fresh Tremella fuciformis with ice cubes in a weight ratio of 1:18-22, and subjecting the mixture to microwave radiation treatment, wherein the microwave radiation intensity of the microwave radiation treatment is 4000-5000 MHz and the treatment time is 3-5 minutes, thereby obtaining Tremella fuciformis water;

[0026] Step 3: placing the Tremella water in a bipolar square wave high voltage pulse electric field, wherein the pulse electric field strength is 30-40 kV / cm, the frequency is 1100-1200 Hz, and the treatment time is 2-3 hours, to separate the Tremella from the water, and the obtained water is the extract;

[0027] Step 4: Concentrate the extract to one-fourth to one-fifth of its volume, add 95% ethanol, the volume ratio of ethanol to the extract being 3-4:1, and separate the precipitate, which is the crude Tremella polysaccharide.

[0028] 2. The method for extracting Tremella polysaccharide according to claim 1 is characterized in that, after step 4, it also includes step 5: washing the obtained crude Tremella polysaccharide with anhydrous ethanol, anhydrous ether and acetone in sequence, drying and crushing to obtain Tremella polysaccharide.

[0029] 3. The method for extracting Tremella polysaccharide according to claim 1, characterized in that in step 3, the temperature of the Tremella water is maintained at 30-35°C.

[0030] 4. The method for extracting Tremella polysaccharide according to claim 1, characterized in that in step 1, the temperature of the clean water is 20-23°C and the soaking time is 3-4h.

[0031] 5. The method for extracting Tremella polysaccharide according to claim 1, characterized in that in said step 2, the weight ratio of the quick-frozen fresh Tremella to ice is 1:20, and the microwave radiation intensity of the microwave radiation treatment is 4500 MHz.

[0032] 6. The method for extracting Tremella polysaccharide according to claim 1, characterized in that in the step 3, the pulse electric field strength is 35 kV / cm, the frequency is 1100 Hz, and the processing time is 2.5 h.

[0033] Example 1

[0034] A method for extracting Tremella polysaccharide comprises the following steps:

[0035] Step 1: Wash the fresh Tremella fuciformis, completely immerse it in clean water at 22°C, soak it for 4 hours, and then quickly freeze it;

[0036] Step 2: Mix the quick-frozen fresh Tremella fuciformis with ice cubes in a weight ratio of 1:20, and perform microwave radiation treatment, wherein the microwave radiation intensity of the microwave radiation treatment is 4500 MHz; the treatment time is 4 minutes, and Tremella fuciformis water is obtained;

[0037] Step 3: placing the Tremella water in a bipolar square wave high voltage pulse electric field, wherein the pulse electric field strength is 35 kV / cm, the frequency is 1100 Hz, and the treatment time is 2.5 h, to separate the Tremella from the water, and the obtained water is the extract;

[0038] Step 4: Concentrate the extract to one-fourth to one-fifth of its volume, add 95% ethanol, the volume ratio of ethanol to the extract being 3-4:1, and separate the precipitate, which is the Tremella crude polysaccharide;

[0039] Step 5: The obtained crude Tremella polysaccharide is washed with anhydrous ethanol, anhydrous ether and acetone in sequence, dried and crushed to obtain Tremella polysaccharide.

[0040] Example 2

[0041] A method for extracting Tremella polysaccharide comprises the following steps:

[0042] Step 1: Wash the fresh Tremella fuciformis, completely immerse it in clean water at 10°C for 2 hours, and then quickly freeze the soaked fresh Tremella fuciformis;

[0043] Step 2: Mix the quick-frozen fresh Tremella and ice cubes in a weight ratio of 1:18, and perform microwave radiation treatment, wherein the microwave radiation intensity of the microwave radiation treatment is 4000 MHz; the treatment time is 3 minutes, and Tremella water is obtained;

[0044] Step 3: placing the Tremella water in a bipolar square wave high voltage pulse electric field, wherein the pulse electric field strength is 30 kV / cm, the frequency is 1100 Hz, and the treatment time is 2 h, to separate the Tremella from the water, and the obtained water is the extract;

[0045] Step 4: Concentrate the extract to one-fourth to one-fifth of its volume, add 95% ethanol, the volume ratio of ethanol to the extract being 3:1, and separate the precipitate, which is Tremella crude polysaccharide;

[0046] Step 5: The obtained crude Tremella polysaccharide is washed with anhydrous ethanol, anhydrous ether and acetone in sequence, dried and crushed to obtain Tremella polysaccharide.

[0047] Example 3

[0048] A method for extracting Tremella polysaccharide comprises the following steps:

[0049] Step 1: Wash the fresh Tremella fuciformis, completely immerse it in clean water at 30°C, soak it for 6 hours, and then quickly freeze the soaked fresh Tremella fuciformis;

[0050] Step 2: Mix the quick-frozen fresh Tremella fuciformis with ice cubes in a weight ratio of 1:22, and perform microwave radiation treatment, wherein the microwave radiation intensity of the microwave radiation treatment is 5000 MHz; the treatment time is 5 minutes, and Tremella fuciformis water is obtained;

[0051] Step 3: placing the Tremella water in a bipolar square wave high voltage pulse electric field, wherein the pulse electric field strength is 40 kV / cm, the frequency is 1200 Hz, and the treatment time is 3 hours, to separate the Tremella from the water, and the obtained water is the extract;

[0052] Step 4: Concentrate the extract to one-fourth to one-fifth of its volume, add 95% ethanol, the volume ratio of ethanol to the extract being 4:1, and separate the precipitate, which is the Tremella crude polysaccharide;

[0053] Step 5: The obtained crude Tremella polysaccharide is washed with anhydrous ethanol, anhydrous ether and acetone in sequence, dried and crushed to obtain Tremella polysaccharide.

[0054] Comparative Example 1

[0055] The method for extracting tremella polysaccharide described in Example 1 is different in that step 2 is replaced by mixing quick-frozen fresh tremella with water, and obtaining tremella water after natural thawing.

[0056] Comparative Example 2

[0057] The method for extracting Tremella polysaccharide described in Example 1 is different in that the pulse electric field treatment in step 3 is omitted, and the extraction is performed directly by standing still for 2.5 hours, and then the Tremella is separated from the water, and the obtained water is the extract.

[0058] Comparative Example 3

[0059] The method for extracting Tremella polysaccharide described in Example 1 is different in that, in step 2, the radiation intensity of the microwave treatment is 2000 MHz.

[0060] Comparative Example 4

[0061] The method for extracting Tremella polysaccharide described in Example 1 is different in that, in step 2, the radiation intensity of the microwave treatment is 6000 MHz.

[0062] Comparative Example 5

[0063] The method for extracting Tremella polysaccharide described in Example 1 is different in that, in step 3, the pulse electric field strength is 20 kV / cm and the frequency is 600 Hz.

[0064] Comparative Example 6

[0065] The method for extracting Tremella polysaccharide described in Example 1 is different in that, in step 3, the pulse electric field strength is 60 kV / cm and the frequency is 1600 Hz.

[0066] Experimental example

[0067] The polysaccharide extraction rates of the Tremella polysaccharide extraction methods of Examples 1-3 and Comparative Examples 1-6 were measured using phenol-sulfuric acid colorimetry. The results are shown in Table 1.

[0068] Table 1

[0069] Extraction rate (%) Extraction rate (%) Example 1 62.6 Comparative Example 3 46.2 Example 2 59.3 Comparative Example 4 53.7 Example 3 61.2 Comparative Example 5 42.6 Comparative Example 1 36.2 Comparative Example 6 56.8 Comparative Example 2 32.5

[0070] It can be seen from the results in Table 1 that the tremella polysaccharide extraction rates of the tremella polysaccharide extraction methods involved in Example 1, Example 2, and Example 3 are all higher than those of Comparative Examples 1-6, among which the tremella polysaccharide extraction rate in Example 1 is the highest, reaching 62.6%.

[0071] The above descriptions are merely embodiments of the present invention and are not intended to limit the patent scope of the present invention. Any equivalent transformations made using the contents of the present invention specification, or directly or indirectly applied in related technical fields, are also included in the patent protection scope of the present invention.

Claims

1. A method for extracting Tremella polysaccharide, characterized in that: The following steps are involved: Step 1: Wash the fresh Tremella fuciformis, completely immerse it in clean water at 22°C, soak it for 4 hours, and then quickly freeze it; Step 2: Mix the quick-frozen fresh Tremella fuciformis with ice cubes in a weight ratio of 1:20, and perform microwave radiation treatment, wherein the microwave radiation intensity of the microwave radiation treatment is 4500 MHz; the treatment time is 4 minutes, and Tremella fuciformis water is obtained; Step 3: placing the Tremella water in a bipolar square wave high voltage pulse electric field, wherein the pulse electric field strength is 35 kV / cm, the frequency is 1100 Hz, and the treatment time is 2.5 h, to separate the Tremella from the water, and the obtained water is the extract; Step 4: Concentrate the extract to one-fourth to one-fifth of its volume, add 95% ethanol, the volume ratio of ethanol to the extract being 3-4:1, and separate the precipitate, which is Tremella crude polysaccharide; Step 5: The obtained crude Tremella polysaccharide is washed with anhydrous ethanol, anhydrous ether and acetone in sequence, dried and crushed to obtain Tremella polysaccharide.